| 以油菜脂肪酸延长酶基因fae1为靶标RNAi载体的构建 |
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| 引用本文： | 肖玲,卢长明.以油菜脂肪酸延长酶基因fae1为靶标RNAi载体的构建[J].中国油料作物学报,2004,26(4):6-11. |
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| 作者姓名： | 肖玲 卢长明 |
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| 作者单位： | 中国农业科学院油料作物研究所,农业部遗传改良重点开放实验室,湖北,武汉,430062 |
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| 摘 要： | 通过基因工程手段抑制脂肪酸延长酶基因fae1的表达,以阻止芥酸合成,培育不受高芥酸窜粉影响的低芥酸品种.利用GenBank中的fae1基因序列AF490462为模板设计引物和多聚酶链式反应(PCR)从白菜、甘蓝和甘蓝型油菜的12个不同品种中扩增出长1007bp的fae1基因片段,通过对fae1基因片段DNA序列的测定和序列比较,找到长度为428bp高度保守区作为RNA干涉(RNAi)的靶标区.根据RNA干涉(RNAi)双向表达载体设计原则,将428bp fae1基因片段以正反两个方向插入双向表达载体pMCG161中,两个片段用一个wax基因的内含子连接,植物筛选标记基因采用bar基因,从而构建以油菜fae1基因为靶标的RNAi载体.
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| 关 键 词： | 油菜 载体构建 芥酸 |
| 文章编号： | 1007-9084(2004)04-0006-06 |
Construction of RNAi expression vector targeting the gene fae1 in rapeseeds |
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| XIAO Ling,LU Chang-mingrops Genetic Improvement,Wuhan ,China.Construction of RNAi expression vector targeting the gene fae1 in rapeseeds[J].Chinese Journal of Oil Crop Sciences,2004,26(4):6-11. |
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| Authors: | XIAO Ling LU Chang-mingrops Genetic Improvement Wuhan China |
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| Institution: | XIAO Ling,LU Chang-mingrops Genetic Improvement,Wuhan 430062,China) |
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| Abstract: | By far all the rapeseed varieties with low erucic acid content have been developed by traditional hybridisation method. Such kind of low erucic acid varieties is characterized by elevation of erucic acid content in developing seeds when contaminated by pollens from high erucic acid varieties during flowering stage. This study is designated to block erucic acid biosynthesis by RNAi strategy targeting fatty acid elongase gene fae1 which is a rate-limiting gene for erucic acid synthesis in Brassica. As a result of comparison of fae1 gene sequence among twelve cultivars of B.napus, B.oleracea and B.rapa, a highly conserved sequence of 428bp in length was found as RNAi target site and used in construction of RNAi vector. The fragment with 428bp in length was amplified from fae1 gene of the variety 821,which is a extensively cultivated high erucic acid rapeseed variety and a reverted repeats of this fragment with an intron of wax gene in between was inserted into pGMCG161. Bar gene is the selection marker in this vector. |
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| Keywords: | fae1 PCR RNAi |
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